Competitive DNA transfection formulation via electroporation for human adipose stem cells and mesenchymal stem cells

We developed an optimal nucleofection formulation for human adipose stem cells by using a three-step method that we had developed previously. This method was designed to determine the optimal formulation for nucleofection that was capable of meeting or surpassing the established commercial buffer (Amaxa), in particular for murine adipose stem cells. By using this same buffer, we determined that the same formulation yields optimal transfection efficiency in human mesenchymal stem cells.Our findings suggest that transfection efficiency in human stem cells can be boosted with proper formulation.Cell-based therapies have great potential for the treatment of genetic disorders as well as currently incurable diseases. Stem cells, the most attractive candidate for such therapy, have been tested in the treatment of leukemias [1,2] and in the regrowth of damaged tissue [3]. Adipose-derived stem cells (ASCs) have recently been isolated [4] and characterized [5]. ASCs are a relatively abundant and easily isolated pluripotent cell line, which makes them a promising candidate as a vehicle for stem cell therapy [4,6,7]. ASCs can be modified to differentiate into various cell lineages, including adipogenic, chondrogenic, and osteogenic cell lines [8], as well as into myoblasts and endothelial cells [5]. ASCs have also demonstrated the ability to home to certain types of tumors [9], which makes them a viable option for antitumor cell therapy.In a previous study, we developed a method for optimizing formulations to aid in the delivery of plasmid DNA in the process of nucleofection [10]. Although nucleofection is an effective form of nonviral transfection for many types of stem cells [11], its therapeutic use is limited by the availability of secret formulations developed by the commercial vendor Amaxa, which must be purchased directly from the vendor. Our devised method offers a three-step plan for determining an optimal transfection formulation generated from known chemicals. The us